Pongkitwitoon Benyakan, Sakamoto Seiichi, Nagamitsu Rika, Putalun Waraporn, Tanaka Hiroyuki, Morimoto Satoshi
Department of Pharmaceutical Botany, Faculty of Pharmacy, Mahidol University, Bangkok, Thailand.
Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
Planta Med. 2018 Sep;84(14):1038-1044. doi: 10.1055/a-0578-8689. Epub 2018 Feb 28.
Homoharringtonine (HHT), also known as omacetaxine, is a natural compound found in the genus and is a promising pharmaceutical drug used for the treatment of chronic or accelerated phase chronic myeloid leukemia. As a tool for the quantitative determination of HHT, a specific monoclonal antibody against HHT (MAb 6A1) was generated by conjugates prepared via sodium periodate-mediated oxidation. The developed indirect competitive enzyme-linked immunosorbent assay (icELISA) using MAb 6A1 was found to be highly specific and sensitive with a limit of detection for HHT of 48.8 ng/mL. Validation assays to evaluate precision and accuracy of the method were conducted by the use of intra- and inter-assay analysis, recovery test, and comparison analysis between the amounts of HHT determined by ELISA and high-performance liquid chromatography. These results revealed that the established icELISA using MAb 6A1 is specific, sensitive, and reliable enough to be applied to the qualitative analysis for HHT. Furthermore, the results of this study support the usefulness of sodium periodate as a reagent for the conjugation between alkaloids and proteins for producing specific antibodies.
高三尖杉酯碱（HHT），也被称为奥马西他辛，是一种存在于三尖杉属植物中的天然化合物，是一种有前景的用于治疗慢性或加速期慢性髓性白血病的药物。作为一种定量测定HHT的工具，通过高碘酸钠介导的氧化反应制备的偶联物产生了一种针对HHT的特异性单克隆抗体（单克隆抗体6A1）。使用单克隆抗体6A1开发的间接竞争酶联免疫吸附测定法（icELISA）被发现具有高度特异性和敏感性，HHT的检测限为48.8 ng/mL。通过批内和批间分析、回收率测试以及ELISA法和高效液相色谱法测定的HHT量之间的比较分析，进行了评估该方法精密度和准确性的验证试验。这些结果表明，使用单克隆抗体6A1建立的icELISA特异性强、灵敏度高且可靠，足以应用于HHT的定性分析。此外，本研究结果支持高碘酸钠作为一种用于生物碱与蛋白质偶联以产生特异性抗体的试剂的实用性。
